CN118948705A - An antioxidant composition extracted from camelina plants and its preparation method and application - Google Patents
An antioxidant composition extracted from camelina plants and its preparation method and application Download PDFInfo
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- CN118948705A CN118948705A CN202411218774.1A CN202411218774A CN118948705A CN 118948705 A CN118948705 A CN 118948705A CN 202411218774 A CN202411218774 A CN 202411218774A CN 118948705 A CN118948705 A CN 118948705A
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- camelina sativa
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dermatology (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Birds (AREA)
- Epidemiology (AREA)
- Cosmetics (AREA)
Abstract
The invention discloses a preparation method of an antioxidant composition extracted from camelina sativa plants, which comprises the steps of extracting camelina sativa plants with alcohol, carrying out enzymolysis, carrying out ultrasonic treatment, decoloring, concentrating and freeze-drying to obtain camelina sativa extract I; extracting camelina sativa plants with water, fermenting, performing enzymolysis, performing ultrasonic treatment, performing alcohol precipitation, filtering to obtain filtrate and filter residues, dissolving the filter residues in water, collecting trapped fluid by ultrafiltration, concentrating, freeze-drying to obtain camelina sativa extract II, concentrating the filtrate, injecting the filtrate into an adsorption resin, collecting water-eluted parts, concentrating, freeze-drying to obtain camelina sativa extract III, wherein the former is a composition mainly containing flavonoid compounds, the latter is a saccharide composition with different molecular weights, and the antioxidant composition is compounded by the three components with synergistic effect; the invention also discloses application of the antioxidant composition in preparation of antioxidant cosmetics. The antioxidant composition prepared by the invention has good antioxidant activity, stable property, mildness and no stimulation.
Description
Technical Field
The invention belongs to the technical field of raw materials and preparation of cosmetics, and particularly relates to an antioxidant composition extracted from camelina sativa plants, a preparation method thereof and application thereof in preparation of antioxidant cosmetics.
Background
The natural metabolism of organisms and the influence of external environment can form a large amount of free radicals and Reactive Oxygen Species (ROS), and the excessive free radicals or ROS can cause damage to cell components such as cell walls, lipid membranes, mitochondria, DNA and the like. Normally, the antioxidant system in humans can scavenge these free radicals and ROS, thereby maintaining a balance between oxidation and antioxidant. However, oxidative stress occurs when the human body is unable to scavenge excess free radicals and ROS using the intracellular antioxidant enzyme system and cellular antioxidants, causing a series of skin problems associated with aging. To ameliorate this problem, it is necessary to supplement the body with exogenous antioxidants or to administer substances that aid in the recovery of endogenous antioxidant substances in the body. The traditional antioxidant generally originates from chemical synthesis, has the defects of poor stability, strong toxic and side effects and the like, and the natural antioxidant from plants has the advantages of safety, no toxicity, strong stability, nature and the like, and meets the requirements of consumers. Therefore, the development of natural antioxidants has become a research hotspot in the cosmetic field.
The plant components commonly added in cosmetics comprise plant oil from plant seeds and fruits, and are rich in fat-soluble vitamins and unsaturated fatty acids. The patent with application number CN202110389479.2 discloses an antioxidant and anti-inflammatory composition containing vegetable oil, wherein one or more of avocado oil, linseed oil, perilla seed oil and camellia seed oil are selected to be matched with jojoba oil to be used as an antioxidant mixture, and the oil has oxidation resistance due to easy oxidation, is easy to deteriorate due to factors such as oxygen, light, enzyme and the like, is easy to precipitate and turbid at low temperature, and can generate harmful substances at high temperature. The stronger the antioxidant activity of the plant components, the easier the plant components are oxidized, so that the strong antioxidant activity of the plant extract and the stability of the plant extract in the cosmetic formula become a group of contradictions to be solved.
Flavonoid and polysaccharide extracted from plants have antiinflammatory, antioxidant and antibacterial effects, and can be used as cosmetic raw material. The patent with the application number of CN202410011438.3 discloses a preparation method of a roxburgh rose extract, which adopts a method of adding a surfactant and an extraction solvent to enhance the wetting and solubilization of the surfactant on the cell wall of the roxburgh rose, greatly improves the extraction rate of total flavone and polysaccharide in the roxburgh rose, and ensures that the prepared freeze-dried powder has stable property. However, since the natural components in the plant extraction have the defects of large quantity and complex components, whether the natural components are mixed together or not has a synergistic effect is yet to be studied.
Camelina sativa is an oil-based commercial crop with short growth cycle (85-100 days), strong cold resistance, few plant diseases and insect pests, low nutrition requirement, and suitability for arid, semiarid and other areas. The applicant records in literature 'quality evaluation of nutrition and functional components of camelina sativa plants', that camelina sativa plants contain rich amino acids, polyphenols and flavonoid compounds, and obtains flavonoid extracts by adopting processes such as alcohol extraction and enzymolysis on camelina sativa plants, thus obtaining a patent (application number is CN 202211155824.7). On the basis, the applicant further develops researches around the extraction process and efficacy evaluation of camelina sativa plants, and the synergistic effect among the extracts is represented through experiments, so that a theoretical basis is provided for developing stable and effective antioxidant cosmetics.
Disclosure of Invention
Based on the defects of the prior art, the invention aims to provide an antioxidant composition extracted from camelina sativa plants and a preparation method thereof, wherein the camelina sativa extract I is obtained through alcohol extraction, enzymolysis and post-treatment, the camelina sativa extract II and the camelina sativa extract III are obtained through water extraction, fermentation and enzymolysis in combination and different post-treatments, and the antioxidant composition with good antioxidant activity and stable property is obtained through compounding of the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III.
The invention also discloses application of the antioxidant composition as a cosmetic raw material in preparing antioxidant cosmetics, and the antioxidant composition has better scavenging ability on DPPH free radicals, hydroxyl free radicals and superoxide anion free radicals, is mild and has no stimulation and no toxic or side effect.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
a preparation method of an antioxidant composition extracted from camelina sativa plants comprises the following steps:
(1) Dispersing cleaned, dried and crushed camelina sativa plants in an ethanol water solution, adding a biological compound enzyme I, carrying out enzymolysis for 1-3 hours at 45-65 ℃, carrying out ultrasonic treatment for 0.5-2 hours, and carrying out solid-liquid separation to obtain camelina sativa extract;
(2) Decolorizing, concentrating and freeze-drying the camelina sativa extract obtained in the step (1) to obtain a camelina sativa extract I;
(3) Extracting cleaned, dried and crushed camelina sativa plants with water to obtain camelina sativa water extract; adding bacillus coagulans liquid and saccharomycete liquid for covering membrane of saccharum to the water extract of camelina sativa, and oscillating for 48-72 hours at 25-35 ℃ to obtain camelina sativa fermentation liquor;
(4) Adding biological compound enzyme II into the camelina sativa fermentation liquor obtained in the step (3), carrying out enzymolysis for 2-5 hours at 45-65 ℃, carrying out ultrasonic treatment for 0.5-2 hours, carrying out solid-liquid separation, and concentrating the liquid to obtain extract; adding ethanol into the extract for alcohol precipitation, and filtering to obtain filtrate and filter residue;
(5) Dissolving the filter residue obtained in the step (4) in water, passing through an ultrafiltration membrane with the molecular weight of 1000-3000 Da, collecting trapped fluid, concentrating, and freeze-drying to obtain a camelina sativa extract II; concentrating the filtrate obtained in the step (4), injecting the concentrated filtrate into an adsorption resin, collecting a water eluting part, concentrating, and freeze-drying to obtain a camelina sativa extract III;
(6) Mixing the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III according to the mass ratio of 1-2:3-5:1 to obtain the antioxidant composition.
Preferably, the biological compound enzyme I in the step (1) is formed by mixing protease and pectase according to the mass ratio of 1:2-3, and the adding amount of the biological compound enzyme I is 0.8-2.0% of the mass of the camelina sativa plant in the step (1); the biological compound enzyme II in the step (4) is formed by mixing cellulase and pectase according to the mass ratio of 1:1.2-2.5, and the addition amount of the biological compound enzyme II is 1.5-3.0% of the mass of the camelina sativa plant in the step (3).
Preferably, the volume concentration of the ethanol in the ethanol water solution in the step (1) is 60-80%; when the camelina sativa plants are dispersed in the ethanol water solution, the dosage of the camelina sativa plants and the ethanol water solution is 1g to 20-40 mL according to the feed-liquid ratio.
Preferably, in the step (2), a decoloring agent is added into the camelina sativa extract, and decoloring is carried out for 1-2 hours at 50-70 ℃; wherein the decoloring agent is formed by mixing diatomite and active carbon according to the mass ratio of 1:1-3, and the ratio of the adding amount of the decoloring agent to the mass of the camelina sativa plant in the step (1) is 1:10-30.
Preferably, the specific step of extracting with water in the step (3) is as follows: dispersing the cleaned, dried and crushed camelina sativa plants in water, and extracting for 1-3 times under normal pressure and a closed environment; controlling the temperature to be 85-100 ℃ and the duration to be 1-3 hours during each extraction, cooling to room temperature after each extraction, filtering out water extract, and combining the water extract extracted each time to obtain camelina sativa water extract; wherein the total amount of the extraction water is 8-30 times of the mass of the camelina sativa plants.
Further, in order to ensure the extraction effect and improve the extraction efficiency, the specific steps of the extraction in the step (3) are as follows: dispersing camelina sativa plants in water (water is used as an extraction solvent), heating to 85-100 ℃ under normal pressure and a closed environment, stirring and extracting for 1-3 h, cooling to room temperature, filtering to obtain primary water extract and slag, and finishing primary extraction; adding water into the slag again, heating to 85-100 ℃ under normal pressure and closed environment, stirring and extracting for 1-2 h, cooling to room temperature, filtering to obtain secondary water extract, and finishing secondary extraction; combining the primary water extract and the secondary water extract to obtain a camelina sativa water extract, and sterilizing for later use; wherein, the water (extraction solvent) consumption is 8-15 times of the camelina sativa plant quality in the primary extraction, and the water (extraction solvent) consumption is 4-8 times of the camelina sativa plant quality in the secondary extraction.
Preferably, in the step (3), the concentration of the bacillus coagulans liquid and the sacculus laminating film saccharomycetes liquid is 0.7-1.3X10 9 cfu/mL, the volume ratio of the bacillus coagulans liquid to the sacculus laminating film saccharomycetes liquid is 1:0.8-1.2, and the total volume of the bacillus coagulans liquid and the sacculus laminating film saccharomycetes liquid is 3-6% of the volume of the camelina sativa water extract.
Further, the bacillus coagulans liquid and the saccharomycete liquid for covering the sacculus film are prepared by the following steps: respectively taking a bacillus coagulans strain and a sacculus tectorial membrane spore yeast strain as strains to be activated, adopting sterile water or a liquid culture medium as a diluent, diluting the strains to be activated by the diluent, inoculating the strains to a solid culture medium, culturing for 48-72 hours at the temperature of 27-35 ℃, growing single bacterial colonies, transferring the single bacterial colonies into the liquid culture medium, sealing by using a breathable film, and performing shake culture for 22-30 hours at the temperature of 25-35 ℃, and then regulating the concentration of bacterial liquid to 0.7-1.3X10 9 cfu/mL to obtain bacillus coagulans liquid and sacculus tectorial membrane spore yeast liquid respectively; wherein, when the strain to be activated is diluted by a diluent, the dosage of the strain to be activated and the diluent is 0.1-0.5 g/mL; when the single colony is transferred to the liquid culture medium, the dosage of the single colony and the YM liquid culture medium is 0.05-0.25 g/L.
Wherein, the solid culture medium adopts YM solid culture medium, and the preparation method of the YM solid culture medium is as follows: taking 1L of dosage as a reference, weighing 4-5 g of peptone, 5-8 g of glucose, 5-10 g of beef extract, 2.5-3 g of yeast extract powder, 16-20 g of agar powder and 4-5 g of sodium chloride, uniformly mixing, adding water to supplement 1L, sterilizing, pouring into a culture dish, standing at room temperature, and solidifying to obtain the YM solid culture medium; the liquid culture medium adopts YM liquid culture medium, and the preparation method of the YM liquid culture medium comprises the following steps: taking 1L of dosage as a reference, weighing 4-5 g of peptone, 5-8 g of glucose, 5-10 g of beef extract, 2.5-3 g of yeast extract powder and 4-5 g of sodium chloride, uniformly mixing, adding water to complement 1L, and sterilizing to obtain the YM liquid culture medium.
Preferably, in steps (2) and (5), the conditions of freeze drying are: vacuum degree is 20-80 Pa, temperature is-20 to-40 ℃ and duration is 3-6 h.
Preferably, in the step (4), ethanol is added into the extract, and the mixture is left for 16 to 36 hours; wherein the addition amount of the ethanol is 1.0 to 2.5 times of the mass of the extract.
Preferably, in the step (5), the adsorption resin adopts D101 macroporous resin, and the dosage of the D101 macroporous resin is 1.5-3 times of the mass of the camelina sativa plant in the step (3); during elution, water is adopted as an eluent, the dosage of the eluent is 4-6 times of the mass of the camelina sativa plants in the step (3), and the eluting speed is 1.5-2.5 Bv/h.
The antioxidant composition prepared by the method is applied to the preparation of antioxidant cosmetics. The invention also provides an antioxidant cosmetic, which comprises the antioxidant composition.
According to the prior art, most of the technologies for researching the Guan Yama camelina sativa plants are content measurement, volatile component measurement and planting technologies, and related documents about the application of camelina sativa plant extracts as raw materials in cosmetics are not available. The invention takes renewable camelina sativa plants as raw materials, and utilizes flavone products and saccharide products generated by fermentation and enzymolysis technologies, thereby reducing the influence of the cosmetic industry on the environment, improving the biological activity of the raw materials, and ensuring that the raw materials have stronger antioxidant function and stable property; the antioxidant composition prepared by mixing three extracts of camelina sativa has better scavenging ability on DPPH free radical, hydroxyl free radical and superoxide anion free radical.
Compared with the prior art, the invention has the following beneficial effects:
The camelina sativa extract I is a composition mainly containing flavonoid compounds, the camelina sativa extract II and the camelina sativa extract III are saccharide compositions with different molecular weights, and the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III have certain scavenging capacity on DPPH free radicals, hydroxyl free radicals and superoxide anion free radicals. The monosaccharide composition, the functional group, the configuration and the microscopic morphology of the saccharide compositions with different molecular weights are different, so that the biological activities of the saccharide compositions are different, and the camelina sativa extract II and the camelina sativa extract III are compounded together and can be mutually modified to achieve the optimization effect; when the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III are mixed and enter a solution system, the camelina sativa extract II and the camelina sativa extract III are used as natural macromolecular polymers and can be combined with the camelina sativa extract I in a bonding and physical adsorption mode, so that the camelina sativa extract II and the camelina sativa extract III become carriers or coating layers of the camelina sativa extract I, the problems of poor water solubility and low chemical stability of the camelina sativa extract I are solved, and the three are compounded to generate obvious synergistic effect in an antioxidant activity test. The antioxidant composition prepared by the invention has the advantages of easy acquisition, good stability, good biocompatibility, no toxicity, no stimulation and the like, shows good antioxidant property, and has good application prospect in preparing antioxidant cosmetics.
Drawings
FIG. 1 is a graph showing the DPPH clearance of the test samples of examples 1 to 3 and comparative examples 7 and 10;
FIG. 2 is a graph showing the superoxide radical scavenging rate of the test samples of examples 1 to 3 and comparative examples 7 and 10;
FIG. 3 is a graph showing the radical scavenging rate of hydroxyl groups of the test samples of examples 1 to 3 and comparative examples 7 and 10.
Detailed Description
In order to make the technical objects, technical solutions and advantageous effects of the present invention more apparent, the technical solutions of the present invention will be further described with reference to specific examples, which are intended to illustrate the present invention but are not to be construed as limiting the present invention, and specific techniques or conditions are not specified in the examples, and are performed according to techniques or conditions described in the literature in the art or according to the product specifications.
The instruments and materials used in the examples below are all commercially available products. The instrument parts used are listed below: ME204 one ten thousandth electronic balance (Metrehler Tolyduo, shanghai), AUW220D one ten thousandth electronic balance (Shimadzu, japan), pipette gun: 100 μL, 200 μL, 1000 μL (Eppendorf), FW-80 high-speed universal pulverizer (Beijing Yongguang), HH-6 digital display constant temperature water bath (He Zhi Bo Rui instruments Co., ltd.), freeze dryer LGJ-12 (Beijing pine source Hua Xing), ultrasonic extractor KQ-500E (Kunshan ultrasonic instruments Co., ltd.), ultraviolet spectrophotometer TU1810 (Beijing Pu analysis); the material fractions used are listed below: camelina sativa plants (obtained from henna Tangyin), aschersonia aleyrodis (strain No. cic 33190), bacillus coagulans (strain No. cic 23843), lactobacillus plantarum (strain No. cic 20272), lactobacillus acidophilus (strain No. cic 20985) and saccharomycetes (strain No. cic 1001) are all purchased from the chinese industrial microbiological culture collection center, ethanol, sodium chloride and the like are all commercially available analytical pure, cellulase (20000U/g) and pectinase (20000U/g) are purchased from shanghai-derived leaf biotechnology company, and protease (100000U/g) is purchased from Xia Cheng (beijing) biotechnology development company.
In the following examples, the bacillus coagulans solution and the saccharomycete solution for covering the sacculus are prepared by the following steps: the method comprises the steps of taking two strains of bacillus coagulans and saccharomycetes of covered with a film at the temperature of 4 ℃ and preserving in a refrigerator inclined surface test tube, respectively adhering 0.03g of bacteria on the surface layer of a bacterial colony on the inclined surface of the two strains by a light spot, respectively diluting and dispersing the bacteria by a 100 mu LYM liquid culture medium, respectively coating the bacteria on a YM solid culture medium by a coating rod, culturing the bacteria for 48 hours at the temperature of 30 ℃, growing single bacterial colony, transferring 0.03g of single bacterial colony into a 200mLYM liquid culture medium, culturing the bacteria for 24 hours (shaking table rotating speed of 180 rpm) at the temperature of 30 ℃, and regulating the concentration of bacterial liquid to be 1.0x10 9 cfu/mL by using a proper amount of YM liquid culture medium to obtain bacillus coagulans liquid and saccharomycetes of covered with film.
The preparation method of the YM solid culture medium comprises the following steps: weighing 4.5g of peptone, 6g of glucose, 6g of beef extract, 3g of yeast extract powder, 18g of agar powder and 4.5g of sodium chloride by taking 1L of dosage as a reference, adding the weighed materials into a 2L triangular flask for mixing, adding water to complement 1L after mixing uniformly, sterilizing at the temperature of 121 ℃ for 20min, pouring into a 90mm culture dish after sterilization, and standing and solidifying at room temperature to obtain the YM solid culture medium; the preparation method of the YM liquid culture medium comprises the following steps: taking 1L of dosage as a reference, weighing 4.5g of peptone, 6g of glucose, 6g of beef extract, 3g of yeast extract powder and 4.5g of sodium chloride, adding the weighed materials into a 2L triangular flask for mixing, adding water to complement 1L after mixing uniformly, and then sterilizing at the temperature of 121 ℃ for 20min to obtain the YM liquid culture medium.
In the following examples, the total flavone content measurement was performed by referring to the total flavone measurement method under the first haw leaf extract item of the year 2020 edition of chinese pharmacopoeia, and the total sugar content measurement was performed by referring to the total sugar measurement method under the first wolfberry item of the year 2020 edition of chinese pharmacopoeia (phenol sulfate method).
The invention provides a preparation method of an antioxidant composition extracted from camelina sativa plants, which comprises the following steps:
(1) Dispersing the cleaned, dried and crushed camelina sativa plant serving as a camelina sativa raw material I in an ethanol water solution, adding a biological compound enzyme I, carrying out enzymolysis for 1-3 hours at 45-65 ℃, carrying out ultrasonic treatment for 0.5-2 hours, and filtering to obtain filter residue I and filtrate I; adding ethanol aqueous solution into filter residue I according to the feed liquid ratio of 1g to 20-30 mL, leaching for 0.5-2 h, filtering to obtain ethanol extract, and combining the ethanol extract and filtrate I to obtain camelina sativa extract;
Wherein the volume concentration of ethanol in the ethanol aqueous solution is 60-80%, and the dosage of the camelina sativa raw material I and the ethanol aqueous solution is 20-40 mL according to the feed-liquid ratio of 1 g; the biological compound enzyme I is formed by mixing protease and pectase according to the mass ratio of 1:2-3, and the addition amount of the biological compound enzyme I is 0.8-2.0% of that of camelina sativa raw material I;
(2) Adding a decoloring agent into the camelina sativa extract obtained in the step (1), decoloring for 1-2 hours at 50-70 ℃, carrying out solid-liquid separation, taking liquid, carrying out rotary evaporation concentration, and carrying out freeze drying to obtain a camelina sativa extract I (the total flavone content is more than 79.2 percent);
Wherein the decoloring agent is formed by mixing diatomite and active carbon according to a mass ratio of 1:1-3, and the ratio of the adding amount of the decoloring agent to the mass of the camelina sativa raw material I is 1:10-30; the conditions of the freeze drying are as follows: vacuum degree is 20-80 Pa, temperature is-20 to-40 ℃ and duration is 3-6 h;
(3) Dispersing the cleaned, dried and crushed camelina sativa plants serving as a camelina sativa raw material II in water, heating the camelina sativa raw material II to 85-100 ℃ under normal pressure and a closed environment, stirring and extracting for 1-3 h, cooling to room temperature, filtering to obtain primary water extract and slag, and finishing primary extraction; adding water into the slag again, heating to 85-100 ℃ under normal pressure and closed environment, stirring and extracting for 1-2 h, cooling to room temperature, filtering to obtain secondary water extract, and finishing secondary extraction; combining the primary water extract and the secondary water extract to obtain a camelina sativa water extract, and sterilizing for later use; adding bacillus coagulans liquid and saccharomycete liquid for covering membrane of saccharum to the water extract of camelina sativa, and oscillating for 48-72 hours at 25-35 ℃ to obtain camelina sativa fermentation liquor;
Wherein, the water consumption is 8-15 times of the mass of the camelina sativa raw material II during primary extraction, and 4-8 times of the mass of the camelina sativa raw material II during secondary extraction; the ratio of the added volumes of the bacillus coagulans liquid and the saccharomycete liquid for covering the cyst is 1:0.8-1.2, and the total volume of the added volumes of the bacillus coagulans liquid and the saccharomycete liquid for covering the cyst is 3-6% of the volume of the water extract of camelina sativa;
(4) Adding biological compound enzyme II into the camelina sativa fermentation liquor obtained in the step (3), carrying out enzymolysis for 2-5 hours at 45-65 ℃, carrying out ultrasonic treatment for 0.5-2 hours, carrying out solid-liquid separation, and concentrating the liquid to obtain extract; adding ethanol into the extract, standing for 16-36 h for alcohol precipitation, and filtering to obtain filtrate II and filter residue II;
The biological compound enzyme II is formed by mixing cellulase and pectase according to the mass ratio of 1:1.2-2.5, and the addition amount of the biological compound enzyme II is 1.5-3.0% of the mass of the camelina sativa raw material II; during alcohol precipitation, the addition amount of the ethanol is 1.0 to 2.5 times of the mass of the extract;
(5) Dissolving the filter residue II obtained in the step (4) in water, passing through an ultrafiltration membrane with a molecular weight of 1000-3000 Da, collecting trapped fluid, concentrating, and freeze-drying to obtain a camelina sativa extract II (with a total sugar content of more than 91.4 percent); concentrating the filtrate II obtained in the step (4), injecting the concentrated filtrate II into an adsorption resin, collecting a water eluting part, concentrating, and freeze-drying to obtain a camelina sativa extract III (total sugar content is more than 90.2%);
Wherein the adsorption resin adopts D101 macroporous resin, and the dosage of the D101 macroporous resin is 1.5-3 times of the mass of the camelina sativa raw material II; during elution, water is adopted as an eluent, the dosage of the eluent is 4-6 times of the mass of the camelina sativa raw material II, and the eluting speed is 1.5-2.5 Bv/h; the conditions of freeze drying are as follows: vacuum degree is 20-80 Pa, temperature is-20 to-40 ℃ and duration is 3-6 h;
(6) Mixing the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III according to the mass ratio of 1-2:3-5:1 to obtain the antioxidant composition.
Example 1
A preparation method of an antioxidant composition extracted from camelina sativa plants comprises the following steps:
(1) Dispersing the cleaned, dried and crushed camelina sativa plants serving as a camelina sativa raw material I in an ethanol water solution, adding a biological compound enzyme I, carrying out enzymolysis for 2.5 hours at 50 ℃, carrying out ultrasonic treatment for 1 hour, and filtering to obtain filter residues I and a filtrate I; adding ethanol water solution into filter residue I according to a feed liquid ratio of 1g to 20mL, leaching for 0.5h, filtering to obtain an ethanol extract, and combining the ethanol extract and filtrate I to obtain a camelina sativa extract;
wherein the volume concentration of ethanol in the ethanol water solution is 75%, and the dosage of the camelina sativa raw material I and the ethanol water solution is 1g to 30mL according to the feed-liquid ratio; the biological compound enzyme I is formed by mixing protease and pectase according to a mass ratio of 1:2, and the addition amount of the biological compound enzyme I is 1% of that of the camelina sativa raw material I;
(2) Adding a decoloring agent into the camelina sativa extract obtained in the step (1), decoloring for 1.5 hours at 60 ℃, carrying out solid-liquid separation, taking liquid, carrying out rotary evaporation and concentration, and carrying out freeze drying to obtain a camelina sativa extract I (total flavone content is 79.8%);
wherein the decoloring agent is formed by mixing diatomite and active carbon according to a mass ratio of 1:2, and the ratio of the adding amount of the decoloring agent to the mass of the camelina sativa raw material I is 1:10; the conditions of the freeze drying are as follows: vacuum degree 30Pa, temperature-30deg.C, and duration 5h;
(3) Dispersing the cleaned, dried and crushed camelina sativa plant serving as a camelina sativa raw material II in water, heating to 90 ℃ under normal pressure and a closed environment, stirring and extracting for 1.5h, cooling to room temperature, filtering to obtain primary water extract and slag, and finishing primary extraction; adding water into the slag again, heating to 90 ℃ under normal pressure and in a closed environment, stirring and extracting for 1h, cooling to room temperature, filtering to obtain secondary water extract, and finishing secondary extraction; mixing the primary water extract and the secondary water extract to obtain water extract of camelina sativa, sterilizing at 121deg.C for 20 min; adding bacillus coagulans liquid and saccharum complex film spore yeast liquid into the camelina sativa water extract, and oscillating for 60 hours at 28 ℃ (the rotation speed of a shaking table is 180 rpm) to obtain camelina sativa fermentation liquor;
Wherein, the water consumption is 12 times of the mass of the camelina sativa raw material II during primary extraction, and 6 times of the mass of the camelina sativa raw material II during secondary extraction; the ratio of the added volumes of the bacillus coagulans liquid and the saccharium knot tectorial membrane saccharomycetes liquid is 1:1, and the added volume of the bacillus coagulans liquid and the saccharium knot tectorial membrane saccharomycetes liquid is 4% of the volume of the camelina sativa water extract;
(4) Adding biological compound enzyme II into the camelina sativa fermentation liquor obtained in the step (3), carrying out enzymolysis for 3 hours at 45 ℃, carrying out ultrasonic treatment for 1 hour, carrying out solid-liquid separation, and concentrating the liquid to obtain extract; adding ethanol into the extract, standing for 28h for alcohol precipitation, and filtering to obtain filtrate II and filter residue II;
The biological compound enzyme II is formed by mixing cellulase and pectinase according to a mass ratio of 1:2, and the addition amount of the biological compound enzyme II is 1.8% of the mass of the camelina sativa raw material II; during alcohol precipitation, the addition amount of the ethanol is 1.5 times of the mass of the extract;
(5) Dissolving the filter residue II obtained in the step (4) in water, passing through an ultrafiltration membrane with a molecular weight of 2000Da, collecting the trapped fluid, concentrating, and freeze-drying to obtain a camelina sativa extract II (total sugar content 91.78%); concentrating the filtrate II obtained in the step (4), injecting into D101 macroporous resin, adsorbing for 1h, taking water as an eluent, collecting water eluting part, concentrating, and freeze-drying to obtain camelina sativa extract III (total sugar content 90.89%);
Wherein the dosage of the D101 macroporous resin is 2 times of the mass of the camelina sativa raw material II, the dosage of the eluent is 5 times of the mass of the camelina sativa raw material II, and the eluting speed is 2Bv/h; the conditions of freeze drying are as follows: vacuum degree 30Pa, temperature-30deg.C, and duration 4h;
(6) Mixing the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 1:3:1 to obtain the antioxidant composition.
Example 2
A preparation method of an antioxidant composition extracted from camelina sativa plants comprises the following steps:
(1) Dispersing the cleaned, dried and crushed camelina sativa plants serving as a camelina sativa raw material I in an ethanol water solution, adding a biological compound enzyme I, carrying out enzymolysis for 3 hours at 50 ℃, carrying out ultrasonic treatment for 1 hour, and filtering to obtain filter residues I and a filtrate I; adding ethanol water solution into filter residue I according to a feed liquid ratio of 1g to 20mL, leaching for 0.5h, filtering to obtain an ethanol extract, and combining the ethanol extract and filtrate I to obtain a camelina sativa extract;
Wherein the volume concentration of ethanol in the ethanol water solution is 70%, and the dosage of the camelina sativa raw material I and the ethanol water solution is 1 g/25 mL according to the feed-liquid ratio; the biological compound enzyme I is formed by mixing protease and pectase according to a mass ratio of 1:3, and the addition amount of the biological compound enzyme I is 1.5% of that of the camelina sativa raw material I;
(2) Adding a decoloring agent into the camelina sativa extract obtained in the step (1), decoloring for 1.5 hours at 60 ℃, carrying out solid-liquid separation, taking liquid, carrying out rotary evaporation and concentration, and carrying out freeze drying to obtain a camelina sativa extract I (the total flavone content is 80.1%);
wherein the decoloring agent is formed by mixing diatomite and active carbon according to a mass ratio of 1:2, and the ratio of the adding amount of the decoloring agent to the mass of the camelina sativa raw material I is 1:20; the conditions of the freeze drying are as follows: vacuum degree 30Pa, temperature-30deg.C, and duration 5h;
(3) Dispersing the cleaned, dried and crushed camelina sativa plant serving as a camelina sativa raw material II in water, heating to 85 ℃ under normal pressure and a closed environment, stirring and extracting for 2 hours, cooling to room temperature, filtering to obtain primary water extract and slag, and finishing primary extraction; adding water into the slag again, heating to 85 ℃ under normal pressure and in a closed environment, stirring and extracting for 1.5h, cooling to room temperature, filtering to obtain secondary water extract, and finishing secondary extraction; combining the primary water extract and the secondary water extract to obtain a camelina sativa water extract, and sterilizing for later use; adding bacillus coagulans liquid and saccharum complex film spore yeast liquid into the camelina sativa water extract, and oscillating for 60 hours at 28 ℃ (the rotation speed of a shaking table is 180 rpm) to obtain camelina sativa fermentation liquor;
Wherein, the water consumption is 10 times of the mass of the camelina sativa raw material II during primary extraction, and 5 times of the mass of the camelina sativa raw material II during secondary extraction; the ratio of the added volumes of the bacillus coagulans liquid and the saccharium knot tectorial membrane saccharomycetes liquid is 1:1, and the added volume of the bacillus coagulans liquid and the saccharium knot tectorial membrane saccharomycetes liquid is 4% of the volume of the camelina sativa water extract;
(4) Adding biological compound enzyme II into the camelina sativa fermentation liquor obtained in the step (3), carrying out enzymolysis for 3 hours at 45 ℃, carrying out ultrasonic treatment for 1 hour, carrying out solid-liquid separation, and concentrating the liquid to obtain extract; adding ethanol into the extract, standing for 28h for alcohol precipitation, and filtering to obtain filtrate II and filter residue II;
The biological compound enzyme II is formed by mixing cellulase and pectinase according to a mass ratio of 1:2, and the addition amount of the biological compound enzyme II is 1.8% of the mass of the camelina sativa raw material II; during alcohol precipitation, the addition amount of the ethanol is 1.5 times of the mass of the extract;
(5) Dissolving the filter residue II obtained in the step (4) in water, passing through an ultrafiltration membrane with a molecular weight of 2000Da, collecting the trapped fluid, concentrating, and freeze-drying to obtain a camelina sativa extract II (total sugar content 91.46%); concentrating the filtrate II obtained in the step (4), injecting into D101 macroporous resin, adsorbing for 1h, taking water as an eluent, collecting water eluting part, concentrating, and freeze-drying to obtain camelina sativa extract III (total sugar content 90.25%);
Wherein the dosage of the D101 macroporous resin is 2 times of the mass of the camelina sativa raw material II, the dosage of the eluent is 5 times of the mass of the camelina sativa raw material II, and the eluting speed is 2Bv/h; the conditions of freeze drying are as follows: vacuum degree 30Pa, temperature-30deg.C, and duration 4h;
(6) Mixing the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 2:5:1 to obtain the antioxidant composition.
Example 3
An antioxidant composition extracted from camelina sativa plants, prepared according to the method of example 1, except for step (6): mixing the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 1.5:4:1.
Comparative example 1
An antioxidant composition extracted from camelina sativa plant is prepared by preparing camelina sativa extract II and camelina sativa extract III according to the method of example 1, and mixing the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 1:1.
Comparative example 2
An antioxidant composition extracted from camelina sativa plant is prepared by preparing camelina sativa extract II and camelina sativa extract III according to the method of example 1, and mixing the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 1:3.
Comparative example 3
An antioxidant composition extracted from camelina sativa plant is prepared by preparing camelina sativa extract II and camelina sativa extract III according to the method of example 1, and mixing the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 2:1.
Comparative example 4
An antioxidant composition extracted from camelina sativa plant is prepared by preparing camelina sativa extract II and camelina sativa extract III according to the method of example 1, and mixing the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 3:1.
Comparative example 5
An antioxidant composition extracted from camelina sativa plant is prepared by preparing camelina sativa extract II and camelina sativa extract III according to the method of example 1, and mixing the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 4:1.
Comparative example 6
An antioxidant composition extracted from camelina sativa plant is prepared by preparing camelina sativa extract II and camelina sativa extract III according to the method of example 1, and mixing the camelina sativa extract II and the camelina sativa extract III according to a mass ratio of 5:1.
Comparative example 7
An antioxidant composition extracted from camelina sativa plant is prepared by the method of example 1, wherein camelina sativa extract I, camelina sativa extract II and camelina sativa extract III are mixed according to a mass ratio of 4:3:1.
Comparative example 8
An antioxidant extract from camelina sativa plants, camelina sativa extract II, was prepared as in example 1.
Comparative example 9
An antioxidant extract from camelina sativa plants, camelina sativa extract III, was prepared as in example 1.
Comparative example 10
An antioxidant extract from camelina sativa plants, camelina sativa extract I, was prepared as in example 1.
Application test
The antioxidant compositions prepared in examples 1 to 3 and comparative examples 1 to 7 and the antioxidant extracts prepared in comparative examples 8 to 9 were used as test samples, and the antioxidant activity, stability and skin irritation of the test samples were measured.
1. Antioxidant Activity assay:
The antioxidation test is to test the scavenging ability of DPPH free radical, hydroxyl free radical and superoxide anion free radical, and the sample preparation method of the antioxidation activity comprises the following steps: 40mg of the sample to be measured was weighed, and 4mL of distilled water was used for dissolution in examples 1 to 3, comparative examples 1 to 6, comparative example 8 and comparative example 9, and since comparative example 7 and comparative example 10 were hardly dissolved in distilled water completely, comparative example 7 and comparative example 10 were dissolved in 4mL of 70% ethanol aqueous solution, and mixed by vortex shaking to prepare a sample mother solution having a final concentration of 10mg/mL, and the sample mother solution was filtered by a filter, diluted with water to a sample solution having a corresponding concentration, and subjected to the next test.
(1) DPPH radical scavenging Capacity determination
Diluting with distilled water respectively, preparing sample solutions with serial concentrations, taking out DPPH reagent from a refrigerator at-20deg.C, balancing at 37deg.C for more than 20min, adding 100mL absolute ethanol into the reagent bottle after the reagent is melted, and shaking vigorously to dissolve the reagent sufficiently to obtain DPPH solution. 1mL of DPPH solution is respectively absorbed, 1mL of absolute ethyl alcohol and 1mL of sample solution with different concentrations are added one by one to be mixed, 1mL of absolute ethyl alcohol and 1mL of sample solution with different concentrations are taken to be mixed one by one, the final solution volume is 2mL, the mixture is sufficiently and uniformly shaken, the mixture is placed at room temperature and kept away from light for 30min to enable the mixture to fully react, then absorbance is measured at 517nm, and vitamin C is used as a positive control sample.
DPPH clearance (%) was calculated according to the following formula:
DPPH clearance (%) = [1- (a 1-A2)/A0 ] ×100;
Wherein A 1 is the absorbance measured after the mixing reaction of the sample solution and the DPPH solution, A 2 is the absorbance measured after the mixing reaction of the sample solution and the absolute ethyl alcohol, and A 0 is the absorbance measured after the mixing reaction of the absolute ethyl alcohol and the DPPH solution.
(2) Determination of the scavenging ability of hydroxyl radicals
Respectively diluting with distilled water to prepare sample solutions with serial concentrations, sequentially adding 50 mu L of sample solution, 50 mu L of salicylic acid solution (9 mmol/L), 50 mu LFeSO 4 of solution (4 mmol/L) and 50 mu LH 2O2 of solution (10 mmol/L) into a 96-well plate to form a sample group, taking deionized water as a blank group instead of the sample solution, and taking deionized water as a control group instead of H 2O2 solution; after 30min of reaction at 37 ℃, absorbance of each group was measured at a wavelength of 510nm, with vitamin C as positive control.
The hydroxyl radical removal (%) was calculated according to the following formula:
Hydroxyl radical clearance (%) = [1- (a Sample of -A Control )/A Blank space ] ×100;
Wherein, A Sample of 、A Control 、A Blank space is the absorbance of the sample group, the control group and the blank group respectively.
(3) Superoxide anion radical scavenging capability test
Respectively diluting with distilled water to prepare sample solutions with serial concentrations, sequentially adding 50 mu L of the sample solution and 100 mu L of Tris-HCl buffer solution into a 96-well plate, shaking, and adding 50 mu L of pyrogallol solution (2 mmol/L) to obtain a sample group; deionized water is used as a control group instead of the pyrogallol solution; deionized water was used instead of the sample solution as a blank, absorbance was measured immediately at 325nm for each group, and vitamin C was used as a positive control sample.
The superoxide radical scavenging (%) was calculated according to the following formula:
Superoxide radical scavenging rate (%) = [1- (a Sample of -A Control )/A Blank space ] ×100;
Wherein, A Sample of 、A Control 、A Blank space is the absorbance of the sample group, the control group and the blank group respectively.
To investigate the synergy between saccharide extracts, test samples of comparative examples 1 to 6 and comparative examples 8 and 9 were first tested, and antioxidant activity data (IC 50) was obtained from the above DPPH clearance, superoxide radical clearance and hydroxyl radical clearance results, as shown in table 1.
Table 1 comparative examples 1 to 6 and comparative examples 8 and 9 were examined for the results of measuring the antioxidant activity of the test samples
Note that: different lower case letters in the same column represent 5% different significance levels, as follows.
As can be seen from table 1, although camelina sativa extract II and camelina sativa extract III both have scavenging ability for DPPH radicals, hydroxyl radicals and superoxide anion radicals, when camelina sativa extract II and camelina sativa extract III are mixed in a mass ratio of 3 to 5:1, the antioxidant activity is significantly better than the others.
Test samples to be tested of examples 1 to 3 and comparative examples 7 and 10 were tested to obtain the DPPH clearance, superoxide radical clearance and hydroxyl radical clearance curves shown in FIGS. 1 to 3, respectively, and antioxidant activity data (IC 50) was obtained from the results of the DPPH clearance, superoxide radical clearance and hydroxyl radical clearance, as shown in Table 2.
Table 2 results of measuring antioxidant activity of test samples of examples 1 to 3 and comparative examples 7 and 10
As can be seen from table 2, the camelina sativa plant extract has better scavenging effect on DPPH free radicals, hydroxyl free radicals and superoxide anion free radicals, the scavenging effect on the free radicals is gradually enhanced along with the increase of the concentration of each component, and a slow down trend is shown at a higher concentration, different components are compounded according to different ratios, the obtained antioxidant activity has obvious difference, wherein the synergistic effect of the compounding of the three extracts is better than that of the compounding of the three components, and the synergistic effect of the compounding of the three components is better than that of the compounding of the three components.
2. Stability test
The antioxidant compositions prepared in examples 1 to 3, the antioxidant extract prepared in comparative example 10 and vitamin C were prepared as solutions having a concentration of 1mg/mL, 50mL of the solutions were placed in an open conical flask, and the mixture was left standing for 6 days in an environment having a temperature of 35℃and a relative humidity of 80% and an illumination intensity of 100Lux, the total sugar and flavone contents of the antioxidant compositions were measured, and the residual ratios were calculated by dividing the measured amounts by the original amounts, and the results were shown in Table 3.
TABLE 3 results of residual Rate measurement
As can be seen from Table 3, the stability of the antioxidant composition prepared according to the present invention is significantly better than that of comparative example 10 and vitamin C.
3. Skin irritation test
16 Healthy adult guinea pigs with intact skin were selected, all of the guinea pigs were shaved 24 hours prior to administration, the hairs on both sides of the back vertebrae of the guinea pigs were carefully removed with a razor, ensuring that no skin damage was caused, and the size of the left and right shaved areas was approximately 2cm x 3cm. 16 guinea pigs were randomly divided into a single administration group and a multiple administration group, 8 in each group, and a physiological saline group and a drug test group were respectively set in the groups.
Prior to administration, the skin was rinsed with warm water and wiped dry with gauze. The antioxidant composition (example 1) was administered to guinea pigs in the single administration group on the right side of the spinal column, and the physiological saline control group was administered on the left side, with the physiological saline group as a negative control. In the experiment, the coating amount of the sample is 0.6g, and the sample is uniformly coated on the shaved part, and sterile gauze is coated on the shaved part and fixed by a medical adhesive tape to prevent the loss of medicines. After the test sample was topically applied to the skin for 4 hours, the covering gauze was removed, and the skin of the applied portion was washed with hot water to remove the remaining test sample. The administration method of guinea pigs in the multiple administration group is the same as that of the single administration group, and the administration is repeated every day for 7 days.
Single skin irritation test: the skin changes (whether erythema and edema phenomenon exists) of the guinea pig application site were visually observed 1h, 24h, 48h, and 72h after the test subjects were removed, and the scores of erythema and edema at each time point were recorded. Multiple skin irritation evaluation: after the test substance is removed 1h, 24h, 48h and 72h from the last administration, the skin change (whether erythema and edema phenomenon exists) of the coated part of the guinea pig is observed with naked eyes, and hair should be sheared if necessary in order to facilitate the coating and result observation of the test substance.
Tests show that for the single administration group and the multiple administration group, the tested object does not cause irritation response of erythema and edema to the skin of the guinea pig when the tested object is the physiological saline and the antioxidant composition, and the average value of the irritation scores is 0 score, so that the single irritation and the multiple irritation are not irritation to the skin.
In summary, the test results of the present invention show that: the camelina sativa plant extract I, the camelina sativa extract II and the camelina sativa extract III and the composition thereof can remove free radicals and have antioxidant activity; meanwhile, when the mass ratio of the camelina sativa plant extract I to the camelina sativa plant extract II to the camelina sativa composition III is 1-2:3-5:1, the antioxidation effect is superior to that of the camelina sativa plant extract I, the camelina sativa plant extract II and the camelina sativa extract III, and a certain synergistic effect is shown. In conclusion, the antioxidant composition prepared by the invention has high antioxidant activity, good stability and no irritation, and can be used as a new cosmetic raw material for preparing antioxidant cosmetics.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (10)
1. A method for preparing an antioxidant composition extracted from camelina sativa plants, which is characterized by comprising the following steps:
(1) Dispersing cleaned, dried and crushed camelina sativa plants in an ethanol water solution, adding a biological compound enzyme I, carrying out enzymolysis for 1-3 hours at 45-65 ℃, carrying out ultrasonic treatment for 0.5-2 hours, and carrying out solid-liquid separation to obtain camelina sativa extract;
(2) Decolorizing, concentrating and freeze-drying the camelina sativa extract obtained in the step (1) to obtain a camelina sativa extract I;
(3) Extracting cleaned, dried and crushed camelina sativa plants with water to obtain camelina sativa water extract; adding bacillus coagulans liquid and saccharomycete liquid for covering membrane of saccharum to the water extract of camelina sativa, and oscillating for 48-72 hours at 25-35 ℃ to obtain camelina sativa fermentation liquor;
(4) Adding biological compound enzyme II into the camelina sativa fermentation liquor obtained in the step (3), carrying out enzymolysis for 2-5 hours at 45-65 ℃, carrying out ultrasonic treatment for 0.5-2 hours, carrying out solid-liquid separation, and concentrating the liquid to obtain extract; adding ethanol into the extract for alcohol precipitation, and filtering to obtain filtrate and filter residue;
(5) Dissolving the filter residue obtained in the step (4) in water, passing through an ultrafiltration membrane with the molecular weight of 1000-3000 Da, collecting trapped fluid, concentrating, and freeze-drying to obtain a camelina sativa extract II; concentrating the filtrate obtained in the step (4), injecting the concentrated filtrate into an adsorption resin, collecting a water eluting part, concentrating, and freeze-drying to obtain a camelina sativa extract III;
(6) Mixing the camelina sativa extract I, the camelina sativa extract II and the camelina sativa extract III according to the mass ratio of 1-2:3-5:1 to obtain the antioxidant composition.
2. The method of preparing an antioxidant composition as claimed in claim 1, wherein: the biological compound enzyme I in the step (1) is formed by mixing protease and pectase according to the mass ratio of 1:2-3, and the adding amount of the biological compound enzyme I is 0.8-2.0% of the mass of the camelina sativa plant in the step (1); the biological compound enzyme II in the step (4) is formed by mixing cellulase and pectase according to the mass ratio of 1:1.2-2.5, and the addition amount of the biological compound enzyme II is 1.5-3.0% of the mass of the camelina sativa plant in the step (3).
3. The method of preparing an antioxidant composition as claimed in claim 1, wherein: the volume concentration of the ethanol in the ethanol water solution in the step (1) is 60-80%; when the camelina sativa plants are dispersed in the ethanol water solution, the dosage of the camelina sativa plants and the ethanol water solution is 1g to 20-40 mL according to the feed-liquid ratio.
4. The method of preparing an antioxidant composition as claimed in claim 1, wherein: in the step (2), a decoloring agent is added into the camelina sativa extract, and decoloring is carried out for 1-2 h at 50-70 ℃; wherein the decoloring agent is formed by mixing diatomite and active carbon according to the mass ratio of 1:1-3, and the ratio of the adding amount of the decoloring agent to the mass of the camelina sativa plant in the step (1) is 1:10-30.
5. The method for preparing an antioxidant composition according to claim 1, wherein the specific step of extracting with water in the step (3) is: dispersing the cleaned, dried and crushed camelina sativa plants in water, and extracting for 1-3 times under normal pressure and a closed environment; controlling the temperature to be 85-100 ℃ and the duration to be 1-3 hours during each extraction, cooling to room temperature after each extraction, filtering out water extract, and combining the water extract extracted each time to obtain camelina sativa water extract; wherein the total amount of the extraction water is 8-30 times of the mass of the camelina sativa plants.
6. The method of preparing an antioxidant composition as claimed in claim 1, wherein: the concentration of the bacillus coagulans liquid and the saccharomycete liquid for covering the sacculus in the step (3) is 0.7-1.3X10 9 cfu/mL, the volume ratio of the bacillus coagulans liquid to the saccharomycete liquid for covering the sacculus is 1:0.8-1.2, and the total volume of the bacillus coagulans liquid and the saccharomycete liquid for covering the sacculus is 3-6% of the volume of the water extract of the camelina sativa.
7. The method for preparing the antioxidant composition according to claim 6, wherein the bacillus coagulans solution and the aschersonia aleyrodis solution are prepared by the following steps: respectively taking a bacillus coagulans strain and a sacculus tectorial membrane spore yeast strain as strains to be activated, adopting sterile water or a liquid culture medium as a diluent, diluting the strains to be activated by the diluent, inoculating the strains to a solid culture medium, culturing for 48-72 hours at the temperature of 27-35 ℃, growing single bacterial colonies, transferring the single bacterial colonies into the liquid culture medium, and after shaking culture for 22-30 hours at the temperature of 25-35 ℃, regulating the concentration of bacterial liquid to be 0.7-1.3X10 9 cfu/mL to obtain bacillus coagulans liquid and sacculus tectorial membrane spore yeast liquid respectively; wherein, when the strain to be activated is diluted by a diluent, the dosage of the strain to be activated and the diluent is 0.1-0.5 g/mL; when the single colony is transferred to the liquid culture medium, the dosage of the single colony and the YM liquid culture medium is 0.05-0.25 g/L.
8. The method of preparing an antioxidant composition as claimed in claim 1, wherein: the conditions of the freeze drying in the steps (2) and (5) are as follows: vacuum degree is 20-80 Pa, temperature is-20 to-40 ℃ and duration is 3-6 h; in the step (4), ethanol is added into the extract, and the mixture is left for 16 to 36 hours; wherein the addition amount of the ethanol is 1.0 to 2.5 times of the mass of the extract; the adsorption resin in the step (5) adopts D101 macroporous resin, and the dosage of the D101 macroporous resin is 1.5-3 times of the mass of the camelina sativa plant in the step (3); during elution, water is adopted as an eluent, the dosage of the eluent is 4-6 times of the mass of the camelina sativa plants in the step (3), and the eluting speed is 1.5-2.5 Bv/h.
9. An antioxidant composition prepared by the method of any one of claims 1 to 8.
10. Use of the antioxidant composition of claim 9 for the preparation of antioxidant cosmetics.
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